LOCUS       sequence-176180-        4692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 558, Emission: 605.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4692)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4692)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
COMMENT     Sequence Label: mRuby-Lifeact-7
FEATURES             Location/Qualifiers
     source          1..4692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        106..210
                     /gene="bla"
                     /label=AmpR promoter
     promoter        212..569
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        212..403
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(235..255)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      420..555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             604..1398
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             604..1398
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(658..677)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1268..1287
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1586..1605)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1630..1677
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2006..2594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2495..2514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2769..3072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3073..3276
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3227..3247
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3305..3314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3311..3361
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             3383..4063
                     /codon_start=1
                     /product="monomeric variant of the red fluorescent protein 
                     eqFP611 (Kredel et al., 2009)"
                     /label=mRuby
                     /translation="MNSLIKENMRMKVVLEGSVNGHQFKCTGEGEGNPYMGTQTMRIKV
                     IEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGVITV
                     MQDTSLEDGCLVYHAQVRGVNFPSNGAVMQKKTKGWEPNTEMMYPADGGLRGYTHMALK
                     VDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVAKFA
                     GLGGG"
     polyA_signal    4188..4309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4225..4244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4279..4298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(4316..4692,1..79))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4453..4474)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4665..4684
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
       61 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      121 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      181 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
      241 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      301 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
      361 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
      421 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
      481 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
      541 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
      601 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
      661 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
      721 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
      781 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
      841 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
      901 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
      961 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     1021 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     1081 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     1141 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     1201 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     1261 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     1321 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     1381 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     1441 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     1501 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     1561 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     1621 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     1681 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     1741 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     1801 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     1861 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1921 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1981 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2041 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2101 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2161 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2221 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2281 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2341 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2401 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2461 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2521 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2581 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2641 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2701 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     2761 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2821 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2881 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2941 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     3001 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     3061 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     3121 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     3181 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     3241 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
     3301 tagcgccacc atgggcgtgg ccgacttgat caagaagttc gagtccatct ccaaggagga
     3361 gggggatcca ccggtcgcca ccatgaacag cctgatcaaa gaaaacatgc ggatgaaggt
     3421 ggtgctggaa ggcagcgtga acggccacca gttcaagtgc accggcgagg gcgagggcaa
     3481 cccctacatg ggcacccaga ccatgcggat caaagtgatc gagggcggac ctctgccctt
     3541 cgccttcgac atcctggcca catccttcat gtacggcagc cggaccttca tcaagtaccc
     3601 caagggcatc cccgatttct tcaagcagag cttccccgag ggcttcacct gggagagagt
     3661 gaccagatac gaggacggcg gcgtgatcac cgtgatgcag gacaccagcc tggaagatgg
     3721 ctgcctggtg taccatgccc aggtcagggg cgtgaatttt cccagcaacg gcgccgtgat
     3781 gcagaagaaa accaagggct gggagcccaa caccgagatg atgtaccccg ctgacggcgg
     3841 actgagaggc tacacccaca tggccctgaa ggtggacggc ggagggcacc tgagctgcag
     3901 cttcgtgacc acctaccgat ccaagaaaac cgtgggcaac atcaagatgc ccggcatcca
     3961 cgccgtggac caccggctgg aaaggctgga agagtccgac aacgagatgt tcgtggtgca
     4021 gcgggagcac gccgtggcca agttcgccgg cctgggcgga gggtaagcgg ccgcgactct
     4081 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4141 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4201 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4261 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     4321 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4381 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4441 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4501 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4561 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4621 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4681 ggagcgggcg ct
//
